中国医科大学学报

中国医科大学学报
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中国医科大学学报 ›› 2018, Vol. 47 ›› Issue (8): 678-681.doi: 10.12007/j.issn.0258-4646.2018.08.002

• 论著 • 上一篇    下一篇

肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域研究

韩旭, 于潜, 田野, 赵希彤, 张磊, 周镝, 田大力   

  1. 中国医科大学附属第四医院胸外科, 沈阳 110032
  • 收稿日期:2018-01-22 出版日期:2018-08-30 发布日期:2018-08-17
  • 通讯作者: 田大力 E-mail:cmu4h_dlt@hotmail.com
  • 作者简介:韩旭(1979-),男,主治医师,博士.
  • 基金资助:
    国家自然科学基金(30973502,30170914);辽宁省教育厅科学研究一般项目(L2013302)

Interacting Domain between ABCE1-encoding Protein and the Cytoskeleton Protein Actin

HAN Xu, YU Qian, TIAN Ye, ZHAO Xitong, ZHANG Lei, ZHOU Di, TIAN Dali   

  1. Department of Thoracic Surgery, The Fourth Affiliated Hospital, China Medical University, Shenyang 110032, China
  • Received:2018-01-22 Online:2018-08-30 Published:2018-08-17

摘要: 目的 明确肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域。方法 通过转化技术,在E.coli BL21(DE3)中诱导表达前期研究获得的表达载体pGEX-4T-1-ABCE1-55,获得GST-ABCE1-55重组融合蛋白表达,应用GST pull-down的方法验证pGEX-4T-1-ABCE1-55与β-肌动蛋白有无相互作用。结果 GST-ABCE1-55是带有GST标签的含有357个氨基酸的重组蛋白,大小约66×103,在GST pull-down实验中,GST-ABCE1-55并不能与β-肌动蛋白特异结合。结论 肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域应位于ABCE1蛋白第358位至第600位编码氨基酸中。

关键词: ATP结合盒转运子E1, pGEX-4T-1原核表达载体, 重组蛋白表达, 蛋白质相互作用结构域

Abstract: Objective To investigate the interacting domain between ABCE1-encoding protein and the cytoskeleton protein actin. Methods Escherichia coli cells were induced with IPTG to produce the recombinant protein glutathione S-transferase(GST)-ABCE1-55 using the prokaryotic expression vector pGEX-4T-1-ABCE1-55,which was previously acquired. A GST pull-down assay was conducted to verify the interaction between GST-ABCE1-55 and β-actin. Results The recombinant protein GST-ABCE1-55 contained a GST tag and was 66×103. There was no interaction between GST-ABCE1-55 and β-actin. Conclusion The interacting domain was predicted to be present between the 358th and 600th encoded amino acids.

Key words: ATP-binding cassette E1, prokaryotic expression vector pGEX-4T-1-ABCE1, recombinant protein expression, interacting domain

中图分类号: 

  • R734.2
[1] BISBAL C,MARTINAND C,SILHOL M,et al.Cloning and characterization of a RNase L inhibitor.A new component of the interferon-regulated 2-5 A pathway[J].Biol Chem,1995,270(22):13308-13317.DOI:10.1074/jbc.270.22.13308.
[2] LE ROY F,BISBAL C,SILHOL M,et al.The 2-5A/RNase L/RNase L inhibitor (RLI)[correction of (RNI)]pathway regulates mitochondrial mRNAs stability in interferon alpha-treated H9 cells[J].Biol Chem,2001,276(51):48473-48482.DOI:10.1074/jbc.M107482200.
[3] BARTHELME D,DINKELAKER S,ALBERS SV,et al.Ribosome recycling depends on a mechanistic link between the FeS cluster domain and a conformational switch of the twin-ATPase ABCE1[J].PNAS,2011,108(8):3228-3233.DOI:10.1073/pnas.1015953108.
[4] PISAREVA VP,SKABKIN MA,HELLEN CU,et al.Dissociation by Pelota,Hbs1 and ABCE1 of mammalian vacant 80S ribosomes and stalled elongation complexes[J].EMBO J,2011,30(9):1804-1817.DOI:10.1038/emboj.2011.93.
[5] ZIMMERMAN C,KLEIN KC,KISER PK,et al.Identification of a host protein essential for assembly of immature HIV-1 capsids[J].Nature,2002,415(6867):88-92.DOI:10.1038/415088a.
[6] LINGAPPA JG,DOOHER JE,NEWMAN MA,et al.Basic residues in the nucleocapsid domain of Gag are required for interaction of HIV-1 Gag with ABCE1(HP68),a cellular protein important for HIV-1 capsid assembly[J].J Biol Chem,2006,281(7):3773-3784.DOI:10.1074/jbc.M507255200.
[7] 刘大治,田大力,任翼.ABCE1在人肺腺癌组织中的表达及其临床意义[J].中华肿瘤杂志,2008,30(4):296-297.
[8] HUANG B,GAO Y,TIAN DL.A small interfering ABCE1-targeting RNA inhibits the proliferation and invasiveness of small cell lung cancer[J].Inter J Mole Med,2010,25(5):687-693.DOI:org/10.3892/ijmm_00000392.
[9] 高英,徐慧慧,王睿,等.新转移相关基因ATP结合盒转运子在非小细胞肺癌中的表达及意义[J].中国医科大学学报,2011,40(10):911-914.
[10] REN Y,LI Y,TIAN DL.Role of the ABCE1 gene in human lung adenocarcinoma[J].Oncol Rep,2012,27(4):965-970.DOI:10.3892/or.2012.1646.
[11] TIAN Y,TIAN X,HAN X,et al.Expression of ATP binding cassette E1 enhances viability and invasiveness of lung adenocarcinoma cells in vitro[J].Mol Med Rep,2016,14(2):1345-1350.DOI:10.3892/mmr.2016.5388.
[12] TIAN Y,HAN X,TIAN DL.The biological regulation of ABCE1[J].IUBMB Life,2012,64(10):795-800.DOI:10.1002/iub.1071.
[13] HAN X,TIAN Y,TIAN DL.Tumor metastatic promoter ABCE1 interacts with the cytoskeleton protein actin and increases cell motility[J].Oncol Reports,2016,35(6):3623-3629.DOI:10.3892/or.2016.4751.
[14] 韩旭,田野,田大力.转移相关基因ABCE1编码蛋白的表达及鉴定[J].生物医学工程与临床,2017,21(3):315-320.
[15] HEIMERL S,BOSSERHOFF AK,LANQMANN T,et al.Mapping ATP-binding cassette transporter gene expression profiles in melanocytes and melanoma cells[J].Melanoma Res,2007,17(5):265-273.DOI:10.1097/CMR.0b013e3282a7e0b9.
[16] HENDIG D,LANQMANN T,ZARBOCK R,et al.Characterization of the ATP-binding cassette transporter gene expression profile in Y79:a retinoblastoma cell line[J].Mol Cell Biochem,2009,328(1/2):85-92.DOI:10.1007/s11010-009-0077-6.
[17] HLAVATA I,MOHELNIKOVA-DUCHONOVA B,VACLAVIKOVA R,et al.The role of ABC transporters in progression and clinical outcome of colorectal cancer[J].Mutagenesis,2012,27(2):187-196.DOI:10.1093/mutage/ger075.
[18] HUANG B,ZHOU H,LANG X,et al.siRNA-induced ABCE1 silencing inhibits proliferation and invasion of breast cancer cells[J].Mol Med Rep,2014,7(2):584-592.DOI:10.3892/mmr.2014.2424.
[19] HLAVAC V,BRYNYCHOVA V,VACLAVIKOVA R,et al.The expression profile of ATP-binding cassette transporter genes in breast carcinoma[J].Pharmacogenomics,2013,14(5):515-529.DOI:10.2217/pgs.13.26.
[20] HUANG B,GONG X,ZHOU H,et al.Depleting ABCE1 expression induces apoptosis and inhibits the ability of proliferation and migration of human esophageal carcinoma cells[J].Int J Clin Exp Pathol, 2014,7(2):584-592.
[21] SHICHIJO,S,ISHIHARA Y,AZUMA K,et al.ABCE1,a member of ATP-binding cassette transporter and tumor-reactive cytotoxic T lymphocytes in colon cancer patients[J].Oncol Rep,2005,13(5):907-913.DOI:org/10.3892/or.13.5.907.
[22] FURUTA M,KOZAKI KI,TANAKA S,et al.miR-124 and miR-203 are epigenetically silenced tumor-suppressive microRNAs in hepatocellular carcinoma[J].Carcinogenesis,2010,31(5):766-776.DOI:10.1093/carcin/bgp250.
[23] SHEA PR,ISHWAD CS,BUNKER CH,et al.RNASEL and RNASEL-inhibitor variation and prostate cancer risk in Afro-Caribbeans[J].Prostate,2008,68(4):354-359.DOI:10.1002/pros.20687.
[24] FIELDS S,SONG O.A novel genetic system to detect protein-protein interactions[J].Nature,1989,340(6230):245-246.DOI:10.1038/340245a0.
[25] CHIEN CT,BATEL PL,STERNGLANZ R,et al.The two-hybrid system:a ethod to identify and clone genes for proteins that interact with a protein of interest[J].Proc Natl Acad Sci USA,1991,88(21):9578-9582.DOI:org/10.1073/pnas.88.21.9578.
[26] KARCHER A,BUTTNER K,MARTENS B,et al.X-ray structure of RLI,an essential twin cassette ABC ATPase involved in ribosome biogenesis and HIV capsid assembly[J].Structure,2005,13(4):649-659.DOI:10.1016/j.str.2005.02.008.
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